IJFS#1599_bozza Ital. J. Food Sci., vol. 32, 2020 - 645 PAPER FATTY ACID PROFILE AND SENSORY PROPERTIES OF ROE DEER MEAT AFTER MODIFIED ATMOSPHERE STORAGE T. DASZKIEWICZ* and J. KONDRATOWICZ Department of Commodity Science and Animal Raw Material Processing, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-957 Olsztyn, Poland *Corresponding author: tomasz.daszkiewicz@uwm.edu.pl ABSTRACT The aim of this study was to determine the effect of cold storage (2°C, 7 and 21 days) under vacuum and modified atmosphere (MA) conditions on the fatty acid (FA) profile and sensory properties of the Longissimus thoracis et lumborum from male roe deer. The total content of polyunsaturated FA tended to decrease during storage. Storage conditions had a limited (P>0.05) influence on the FA composition. Vacuum- and MA-packaged meat was characterized by high sensory quality during storage. However, samples packaged in MA composed of 40% CO2+60% N2 had a tendency to higher average scores for taste desirability after the third week of storage. Keywords: game, cold storage, meat quality Ital. J. Food Sci., vol. 32, 2020 - 646 1. INTRODUCTION The consumption of game meat is relatively low despite its high nutritional and culinary value (SCHULP et al., 2014). There are various reasons for the above, and one of them is a low variety of venison preservation techniques on the consumer market. In retail, game meat is available mostly in the form of vacuum-packaged and deep frozen products. Vacuum packaging inhibits the growth of aerobic microorganisms, limits lipid oxidation and prolongs the product's shelf life (STELLA et al., 2018), but it also has several drawbacks, such as the dark color of meat and considerable drip loss, which compromises consumer acceptance (SAKOWSKA et al., 2016). The meat of wild animals is vacuum packaged on account of its unique quality attributes. Game meat is characterized by a high content of unsaturated phospholipids (VALENCAK et al., 2015) and heme iron in myoglobin (WIKLUND et al., 2006), and it is susceptible to auto-oxidation, which compromises its quality. To preserve the freshness of venison, oxygen is removed from the packaging during the vacuum packaging process. Freezing decreases water activity, slows down chemical and biochemical processes, inhibits microbial growth and extends the product's shelf life (ZHOU et al., 2010). However, freezing, frozen storage and thawing lead to adverse changes in meat quality. Modified atmosphere (MA) packaging preserves the attributes of fresh meat (DJENANE and RONCALÉS, 2018). The gas composition of MA is selected for a given type of meat to prevent undesirable changes in quality caused by microbial growth and oxidation or to preserve its attractive color (beef) (ZHANG et al., 2015). Modified atmosphere packaging eliminates the drawbacks of vacuum packaging, such as considerable drip loss as well as packaging and product deformation, through the application of reduced pressure that increases consumer acceptance (ŠČETAR et al., 2010). The effects of storage conditions (MA composition) and cold storage time on the quality of meat from farm animals, including poultry, and fish have been extensively researched (ZHANG et al., 2015). However, their influence on the quality of venison, in particular meat from wild-living animals, remains insufficiently investigated. Therefore, the objective of this study was to determine the effect of vacuum packaging and MA packaging on the fatty acid profile and sensory properties of meat from male roe deer (Capreolus capreolus L.). 2. MATERIALS AND METHODS 2.1. Sampling, packaging and storage The experimental materials comprised samples of the Longissimus thoracis et lumborum (LTL) muscle collected from the carcasses of 16 male roe deer aged 3 to 5 years, hunter- harvested in the forests of North-Eastern Poland (Sępopol Plain, Region of Warmia and Mazury) in June and July during one hunting season. During carcass dressing in a meat processing plant (within 48-54 h of harvest), right and left LTL muscles were cut out, placed in polyethylene bags on ice, and transported to the laboratory. Each muscle was divided into samples of similar weight, which were allocated to groups A, B, C and D. Samples A were immediately subjected to laboratory analyses, samples B were vacuum- packaged, samples C and D were packaged in MA containing 40% CO2+60% N2 and 60% CO2+40% N2, respectively. The samples were packaged in barrier bags made of ethylene- vinyl alcohol (EVOH) copolymer with the following gas permeability: O2=1 ml/ m2/24 h/bar/23°C, N2<0.1 ml/m2/24 h/bar/23°C, CO2=1.6 ml/m2/24 h/bar/23°C, H2O=3 Ital. J. Food Sci., vol. 32, 2020 - 647 g/m2/24 h/23ºC using the PP-5MG (015) vacuum packaging machine (TEPRO S.A., Koszalin, Poland). The samples were stored in a cooling chamber without forced air-flow at a temperature of 2ºC for 7 and 21 days. Meat quality was determined based on an analysis of the fatty acid profile of intramuscular fat (IMF), an evaluation of the sensory properties of meat, and shear force measurements. 2.2. Analytical procedures The sensory properties of meat were evaluated after cooking in 0.6% NaCl solution at 96ºC (±2°C) until internal temperature reached 80ºC, as described by DASZKIEWICZ et al. (2012). Five panelists on a 5-point scale (5 points - most desirable, 1 point - least desirable) evaluated meat quality. Prior to the evaluation, the panelists had been trained in the sensory properties of cooked venison based on cooked beef loin as the reference standard. The panelists assessed encoded samples composed of 1 cm x 1 cm x 1 cm meat cubes, cut from the center of each cooked sample, cooled to room temperature. Redistilled water was made available to the panelists for mouth cleansing between samples. The sensory properties (aroma, taste, juiciness, tenderness) of up to 5 meat samples were assessed per session. The maximum shear force required to cut meat samples (5 cylinder-shaped samples, 1.27 cm in diameter, 2 cm in height) across the grain was measured using a Warner-Bratzler head (500 N, speed 100 mm/min) attached to the Instron 5542 universal testing machine (Instron, Canton, Massachusetts, USA). The samples were prepared as described by HONIKEL (1998). Intramuscular fat (IMF) was extracted by Soxhlet extraction (AOAC, 1990) with diethyl ether as the solvent in the Soxtec™ 2050 Auto Fat Extraction System (FOSS Analytical, Hilleroed, Denmark). The fatty acid profile of IMF was determined by gas chromatography using a UNICAM PU-4600 gas chromatograph with a flame ionization detector (FID) on a glass capillary column (length: 2.10 m, inner diameter: 4.0 mm); detector temperature - 250ºC, injector temperature - 225ºC, column temperature - 200ºC, carrier gas - argon, carrier gas flow rate - 50 ml/min. Fatty acid methyl esters were prepared by the modified Peisker method with chloroform: methanol: sulphuric acid (100:100:1 v/v) (ŻEGARSKA et al., 1991). 2.3. Statistical analysis The results were processed statistically by one-way ANOVA with the use of STATISTICA ver. 13.3 software (TIBCO Software Inc.). The statistical significance of differences between mean values in groups was determined using the Bonferroni correction at P≤0.05. 3. RESULTS AND DISCUSSION 3.1. Sensory properties of meat A sensory evaluation confirmed high quality of meat from male roe deer irrespective of storage conditions (Table 1). Stored meat samples were characterized by lower aroma intensity than non-stored samples, and the differences were higher (P≤0.05) in MA- packaged meat. Stored samples received similar scores for aroma intensity, only vacuum- packaged samples stored for 21 days scored higher (P≤0.05) than samples packaged in MA Ital. J. Food Sci., vol. 32, 2020 - 648 containing 60% CO2 and 40% N2. Storage time had no influence (P>0.05) on aroma desirability but after 21 days, MA-packaged meat received higher (P≤0.05) scores for this attribute than vacuum-packaged samples. Table 1. Sensory properties (points) and shear force values (N) of meat from male roe deer stored in a modified atmosphere (arithmetic means±SD). Parameter Storage (days) Vacuum Modified atmosphere 40% CO2+60% N2 60% CO2+40% N2 Aroma - intensity 0 4.13±0.85 4.13±0.85a 4.13±0.85a 7 3.72±0.68 3.38±0.59b 3.28±0.55b 21 3.75±0.86x 3.34±0.54bxy 3.22±0.45by Aroma - desirability 0 5.00±0.00 5.00±0.00 5.00±0.00 7 4.88±0.22 4.94±0.17 4.97±0.13 21 4.87±0.22x 5.00±0.00y 5.00±0.00y Taste - intensity 0 4.09±0.52 4.09±0.52a 4.09±0.52a 7 3.91±0.55 3.78±0.45b 3.75±0.37b 21 4.19±0.31x 4.00±0.00aby 4.06±0.17axy Taste - desirability 0 5.00±0.00a 5.00±0.00a 5.00±0.00a 7 4.84±0.24a 4.84±0.24ab 4.88±0.22a 21 4.28±0.82bx 4.78±0.36by 4.38±0.62bxy Juiciness 0 3.59±0.38 3.59±0.38 3.59±0.38ab 7 3.44±0.54 3.66±0.44 3.69±0.40a 21 3.38±0.43 3.38±0.43 3.38±0.47b Tenderness 0 4.44±0.48a 4.44±0.48a 4.44±0.48 7 4.56±0.57a 4.63±0.50ab 4.72±0.41 21 4.97±0.12bx 4.88±0.29bxy 4.75±0.37y Shear force 0 21.27±4.42a 21.27±4.42a 21.27±4.42a 7 17.98±3.39b 16.94±2.76b 18.86±4.46ab 21 16.24±2.20b 17.14±2.48b 16.72±2.68b Values within a row with different superscript letters (x-y) are significantly different (P≤0.05). Values within a column with different superscript letters (a-b) are significantly different (P≤0.05). During storage, taste intensity decreased in samples stored for 7 days compared with non- stored samples and samples stored for 21 days. This trend was more pronounced (P≤0.05) in MA-packaged meat than in vacuum-packaged meat. An analysis of the effect of storage conditions on taste intensity revealed that after 21 days, vacuum-packaged samples scored higher (P≤0.05) than samples packaged in MA composed of 40% CO2 and 60% N2. Roe deer meat stored for 21 days was characterized by lower (P≤0.05) taste desirability than non-stored meat and meat stored for 7 days. Taste deteriorated at the slowest rate in meat packaged in MA composed of 40% CO2 and 60% N2, and at the fastest rate in vacuum-packaged meat and meat packaged in MA containing 60% CO2 and 40% N2. Similar results were reported by SEMAN et al. (1989) who observed a decrease in the flavor desirability of meat from farm-raised male red deer. In their study, vacuum- packaged samples and samples packaged in MA consisting of 100% CO2 were stored for 6, Ital. J. Food Sci., vol. 32, 2020 - 649 12 and 18 weeks at -1ºC. The undesirable changes in palatability most likely resulted from enzymatic and chemical reactions, and the accumulation of microbial metabolites. When analyzing the effects of chemical processes on the sensory properties of meat, attention should be paid to peroxidation of lipids, in particular polyunsaturated fatty acids (PUFAs) (PAPUC et al., 2017). Meat juiciness was comparable (P>0.05) in non-stored samples and MA-packaged samples stored for 7 days. The value of this attribute decreased after 21 days of storage. Storage conditions had no effect (P>0.05) on meat juiciness. However, MA-packaged samples stored for 7 days received somewhat higher average scores for juiciness than vacuum- packaged samples. No differences (P>0.05) in juiciness were reported by HUR et al. (2013) in beef, PIASKOWSKA et al. (2016) in fallow deer meat and by SEMAN et al. (1989) in red deer meat packaged in MA consisting of CO2 and N2 or 100% CO2 vs. vacuum-packaged samples. ORKUSZ (2018) and CLAUSEN et al. (2009) observed a decrease in the juiciness of samples (goose meat and beef, respectively) stored in MA containing 50-80% O2. The analyzed meat was characterized by desirable tenderness, which further improved during storage, and differences (P≤0.05) were found for vacuum-packaged samples and samples packaged in MA consisting of 40% CO2+60% N2. After 21 days of storage, vacuum-packaged meat had the highest tenderness scores and meat packaged in MA composed of 60% CO2+40% N2 had the lowest tenderness scores (P≤0.05). The tenderness of samples packaged in MA consisting of 40% CO2 and 60% N2 was at an average level. The results of tenderness evaluation were reflected in shear force values, which were lower (P≤0.05) in stored meat than in non-stored samples. No differences (P>0.05) were found between the average shear force values of meat stored for 7 and 21 days. Storage conditions had no effect (P>0.05) on shear force values. The increase in tenderness of roe deer meat and the decrease in shear force values after cold storage, observed in our study, were related to the activity of endogenous and bacterial proteolytic enzymes. According to WIKLUND et al. (2014), meat from selected cervid species does not require aging due to its high tenderness, which probably would not show a further improvement during the process. PAULSEN et al. (2005) found no considerable differences in the average values of shear force between samples of roe deer meat stored in vacuum (3.5ºC, 132 h) and samples collected from roe deer carcasses on day 5 post mortem. SEMAN et al. (1989) noted no differences (P>0.05) in the shear force values of meat from male red deer stored in vacuum and MA (100% CO2) for 18 weeks. 3.2. Fatty acid composition of intramuscular fat Storage time and conditions had no effect (P>0.05) on the content of saturated fatty acids (SFAs) in IMF (Table 2). Nevertheless, an increase in C 16:0 and C 18:0 content (in particular C 18:0) during storage contributed to an increase (P>0.05) in the total SFA pool. Similarly to SFAs, storage time and conditions had a limited influence on the content of UFAs (Table 3). The differences between mean values in groups were small and statistically significant only for margoleic acid (C 17:1) and gadoleic acid (C 20:1). In vacuum-packaged meat, the content of C 20:1 was lower (P≤0.05) in non-stored samples and samples stored for 21 days compared with those stored for 7 days. After 7 days of storage, the content of C 17:1 and C 20:1 was higher (P≤0.05) in vacuum-packaged samples than in samples packaged in MA composed of 60% CO2+40% N2 and 40% CO2+60% N2, respectively. Ital. J. Food Sci., vol. 32, 2020 - 650 Table 2. Percentage of saturated fatty acids in total fatty acids in the intramuscular fat of meat from male roe deer stored under vacuum and modified atmosphere conditions (arithmetic means ± SD). Parameter Storage (days) Vacuum Modified atmosphere 40% CO2+60% N2 60% CO2+40% N2 C 12:0 0 1.06±0.61 1.06±0.61 1.06±0.61 7 1.02±0.49 1.09±0.71 0.92±0.35 21 0.86±0.55 0.98±0.64 0.81±0.49 C 14:0 0 1.96±0.50 1.96±0.50 1.96±0.50 7 1.90±0.27 2.05±0.66 1.81±0.34 21 1.88±0.41 1.97±0.86 1.97±0.45 C 15:0 0 0.57±0.15 0.57±0.15 0.57±0.15 7 0.56±0.18 0.59±0.19 0.53±0.11 21 0.64±0.45 0.59±0.20 0.58±0.14 C 16:0 0 23.72±2.64 23.72±2.64 23.72±2.64 7 23.59±1.47 23.86±1.84 23.58±2.16 21 24.18±1.66 24.39±2.98 24.69±2.84 C 17:0 0 1.37±0.15 1.37±0.15 1.37±0.15 7 1.37±0.15 1.40±0.23 1.35±0.19 21 1.37±0.19 1.39±0.18 1.38±0.19 C 18:0 0 23.91±1.85 23.91±1.85 23.91±1.85 7 24.22±3.14 24.56±2.18 24.03±1.85 21 25.27±2.78 24.63±2.86 25.24±2.32 C 20:0 0 0.44±0.19 0.44±0.19 0.44±0.19 7 0.45±0.17 0.49±0.30 0.39±0.10 21 0.51±0.38 0.41±0.18 0.46±0.26 SFAs 0 53.04±4.13 53.04±4.13 53.04±4.13 7 53.11±3.25 54.04±4.07 52.62±3.48 21 54.72±4.09 54.35±3.78 55.13±3.70 SFAs - saturated fatty acids. Storage time had no effect (P>0.05) on total content of UFAs, but the total content of PUFAs was lower in stored samples, particularly (P≤0.05) those vacuum-packaged and packaged in MA containing 40% CO2 and 60% N2, than in non-stored samples (Table 3). As a result, the PUFAs/SFAs ratio was slightly lower in stored meat (Table 4). The total content of monounsaturated fatty acids (MUFAs) increased (P>0.05) in meat stored for 7 days and decreased (P>0.05) in meat stored for 21 days (Table 3). Increased PUFA content is desirable in view of the nutritional value and health-promoting properties of meat but they can also lead to undesirable changes in the product’s quality during storage. Auto-oxidation of PUFAs contributes to undesirable changes in the aroma and taste of meat as well as to the formation of compounds that decrease the nutritional value of meat, and toxic compounds (PAPUC et al., 2017). Therefore, meat packaging must protect the product against the adverse effects of oxygen. This applies also to the meat of wild animals, which is rich in both PUFAs and heme iron that catalyzes auto-oxidation. In practice, vacuum and MA packaging are used to prevent Ital. J. Food Sci., vol. 32, 2020 - 651 oxygen from entering the package - oxygen is replaced with appropriately selected gas mixtures. Table 3. Percentage of unsaturated fatty acids in total fatty acids in the intramuscular fat of meat from male roe deer stored under vacuum and modified atmosphere conditions (arithmetic means ± SD). Parameter Storage (days) Vacuum Modified atmosphere 40% CO2+60% N2 60% CO2+40% N2 C 14:1 0 0.15±0.08 0.15±0.08 0.15±0.08 7 0.17±0.07 1.12±0.06 0.15±0.07 21 0.41±0.72 0.27±0.42 0.13±0.07 C 16:1 0 2.22±0.20 2.22±0.20 2.22±0.20 7 2.38±0.43 2.50±0.45 2.36±0.38 21 2.28±0.66 2.34±0.38 2.43±0.47 C 17:1 0 0.24±0.04 0.24±0.04 0.24±0.04 7 0.29±0.04x 0.26±0.05xy 0.23±0.02y 21 0.29±0.12 0.27±0.05 0.27±0.10 C 18:1 0 26.28±4.46 26.28±4.46 26.28±4.46 7 27.53±4.38 27.12±4.42 26.54±4.75 21 26.79±3.92 26.02±4.89 25.88±2.99 C 18:2 0 12.07±2.41 12.07±2.41 12.07±2.41 7 11.07±1.95 10.78±1.92 11.94±3.60 21 10.75±2.46 11.39±2.81 11.22±3.25 C 18:3 0 2.19±0.47 2.19±0.47 2.19±0.47 7 1.85±0.53 1.93±0.57 1.98±0.62 21 1.80±0.60 1.92±0.71 1.75±0.59 C 20:1 0 0.39±0.13a 0.39±0.13 0.39±0.13 7 0.55±0.09bx 0.44±0.10y 0.48±0.10xy 21 0.40±0.05a 0.41±0.12 0.42±0.15 C 20:4 0 4,80±4.73 4.80±4.73 4.80±4.73 7 3.06±0.65 2.81±0.77 3.69±1.81 21 2.57±1.25 3.03±1.00 2.77±1.07 MUFAs 0 29.28±4.31 29.28±4.31 29.28±4.31 7 30.92±4.36 30.44±4.23 29.77±4.83 21 30.16±3.46 29.31±4.98 29.13±2.97 PUFAs 0 19.07±4.89a 19.07±4.89a 19.07±4.89 7 15.97±2.74b 15.53±2.83b 17.61±5.80 21 15.12±3.62b 16.35±4.14ab 15.74±4.66 UFAs 0 46.96±4.13 46.96±4.13 46.96±4.13 7 46.89±3.25 45.96±4.07 47.38±3.48 21 45.28±4.09 45.65±3.78 44.87±3.70 MUFAs - monounsaturated fatty acids; PUFAs - polyunsaturated fatty acids; UFAs - unsaturated fatty acids (MUFAs + PUFAs). Values within a row with different superscript letters (x-y) are significantly different (P≤0.05). Values within a column with different superscript letters (a-b) are significantly different (P≤0.05). Ital. J. Food Sci., vol. 32, 2020 - 652 Table 4. The ratio of unsaturated fatty acids to saturated fatty acids in the intramuscular fat of meat from male roe deer stored under vacuum and modified atmosphere conditions (arithmetic means ± SD). Parameter Storage (days) Vacuum Modified atmosphere 40% CO2+60% N2 60% CO2+40% N2 UFAs/SFAs 0 0.90±0.14 0.90±0.14 0.90±0.14 7 0.89±0.11 0.86±0.13 0.91±0.12 21 0.84±0.13 0.85±0.13 0.82±0.12 MUFAs/SFAs 0 0.56±0.11 0.56±0.11 0.56±0.11 7 0.59±0.11 0.57±0.11 0.57±0.11 21 0.56±0.09 0.55±0.12 0.53±0.07 PUFAs/SFAs 0 0.36±0.11a 0.36±0.11a 0.36±0.11 7 0.30±0.05ab 0.29±0.06b 0.34±0.12 21 0.28±0.08b 0.30±0.08ab 0.29±0.10 MUFAs - monounsaturated fatty acids; PUFAs - polyunsaturated fatty acids; UFAs - unsaturated fatty acids (MUFAs + PUFAs). Values within a column with different superscript letters (a-b) are significantly different (P≤0.05). The beneficial effect of oxygen-free packaging that limits lipid peroxidation in game meat was reported by FAROUK and FREKE (2008). They noted a lower content of malondialdehyde (an indicator of auto-oxidative changes) in samples of the Semimembranosus muscle of red deer, which were vacuum-packaged and stored for 9 months, in comparison with samples that were placed in bags that permitted free exchange of air between the inside and outside of the packaging. Unfortunately, vacuum and MA packaging with oxygen-free gas mixtures cannot completely inhibit undesirable changes in the quality of cold-stored meat products. Under industrial conditions, O2 cannot be entirely removed from the vacuum or MA packaging. According to BERRUGA et al. (2005), the presence of “residual oxygen”, less than 2% is sufficient to promote lipid peroxidation. 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